Fas ligand gene transfer to renal allografts in rats: effects on allograft survival.
BACKGROUND
Fas ligand (FasL) induces apoptosis of cells bearing its receptor Fas, and has been shown to be important in T-cell development and regulation and in immune privilege. We hypothesized that FasL expression by renal allografts might provide protection from rejection.


METHODS
The murine FasL cDNA was cloned into a replication-defective adenovirus (AdV-FasL). Protein expression was confirmed by immunostaining of AdV-FasL-transduced HeLa cells. Allogeneic kidney transplants were performed between WF (RT1u) donors and Lewis (RT1) recipients. Donor kidneys were perfused in situ with saline alone (control), or 9 x 10(9) plaque-forming units of AdV-FasL. One native kidney was removed at the time of transplant and the other at 6 or 7 days. Uremic death was the endpoint, and deaths within 7 days of transplant were excluded. Transduced allografts were stained for FasL expression using a monoclonal antibody and tested for FasL mRNA production by reverse transcriptase-polymerase chain reaction and Northern blotting.


RESULTS
Immunostaining of AdV-FasL-transduced allografts demonstrated efficient gene transfer lasting approximately 2 weeks, and FasL mRNA production in the AdV-FasL-transduced allografts was confirmed by Northern blotting and reverse transcriptase-polymerase chain reaction. Mean survival of animals with AdV-FasL-transduced renal allografts was 27.8 days vs. 11.6 days in control animals (P < 0.05).


CONCLUSIONS
(1) Adenoviral vectors can successfully transduce rat kidneys with the FasL cDNA. (2) FasL gene transfer prolongs rat renal allograft survival.
Swenson
K M
KM
Dumont-UCLA Transplant Center, Department of Surgery, UCLA School of Medicine, Cedars-Sinai Medical Center, Los Angeles, California 90095, USA.
Ke
B
B
Wang
T
T
Markowitz
J S
JS
Maggard
M A
MA
Spear
G S
GS
Imagawa
D K
DK
Goss
J A
JA
Busuttil
R W
RW
Seu
P
P
D000256:Adenoviridae; D000818:Animals; D003602:Cytotoxicity, Immunologic / Q000235:genetics; D018076:DNA, Complementary; D053222:Fas Ligand Protein; D018014:Gene Transfer Techniques; D006084:Graft Rejection / Q000276:immunology / Q000378:metabolism* / Q000517:prevention & control; D006085:Graft Survival / Q000235:genetics / Q000276:immunology; D006367:HeLa Cells; D006801:Humans; D007150:Immunohistochemistry; D016030:Kidney Transplantation / Q000276:immunology* / Q000473:pathology; D008562:Membrane Glycoproteins / Q000235:genetics / Q000276:immunology / Q000378:metabolism; D016133:Polymerase Chain Reaction; D012333:RNA, Messenger / Q000032:analysis; D051381:Rats; D011917:Rats, Inbred Lew; D011920:Rats, Inbred WF; D014161:Transduction, Genetic; D014184:Transplantation, Homologous / Q000276:immunology* / Q000473:pathology
D018076:DNA, Complementary; C505661:FASLG protein, human; D053222:Fas Ligand Protein; C505662:Faslg protein, rat; D008562:Membrane Glycoproteins; D012333:RNA, Messenger